Herpes simplex virus (HSV) infection is one of the most prevalent infectious diseases worldwide, with HSV‐2 being primarily associated with genital infections. HSV‐2 is believed to account for the majority of cases of neonatal herpes, which may cause diverse of complications in infected newborns. The present study sought to estimate the prevalence of HSV‐2 in placental tissue samples and the incidence of HSV‐2 in the umbilical cord blood of newborn infants. Placental tissue samples from 201 women (maternal‐side and fetal‐side = 402 specimens) and 184 neonatal cord blood samples, all collected at the obstetric ward of a University hospital were studied. HSV‐2 was detected by means of nested PCR. The prevalence of HSV‐2 in placental samples was 9.0% (n = 18), and the incidence of neonatal HSV‐2 infection was 1.1% (n = 2). All HSV‐2‐positive patients were asymptomatic at the time of delivery and none reported genital herpes. Women with a time between rupture of membranes and delivery of ≥360 min had an approximately fourfold risk of HSV‐2 infection in the placental tissue (95% CI 0.93–5.66, P = 0.01). These results suggest that HSV‐2 is present in the placenta of asymptomatic women and that a risk of transmission to the neonate exists. New strategies must be implemented for the management of asymptomatic patients who are capable of transmitting the virus to the newborn. J. Med. Virol. 86:519–524, 2014. © 2013 Wiley Periodicals, Inc.